Intranasal administration

ABSTRACT

Intranasal administration of proteins, such as insulin and insulin analogues, in particular immunogenic proteins to the upper posterior region of a nasal cavity of a subject, and in particular the olfactory bulb region.

The present invention relates to intranasal administration of proteins,such as insulin and insulin analogues, in particular immunogenicproteins to the upper posterior region of a nasal cavity of a subject,and in particular the olfactory bulb region.

Recent studies have demonstrated that the intranasal administration ofinsulin acts to improve memory and has an effect on obesity [1-8].

However, other studies have shown that intranasal administration ofinsulin over an extended period, typically six months, results inantibodies being raised to insulin [9].

Whilst the authors of this publication argue that the raising ofantibodies to insulin could be beneficial in terms of being protectiveof loss of beta cell function in individuals at risk of Type I diabetes,there is no recognition that raising a neutralising antibody response toinsulin would be of particular concern in patients with obesity who maybe at risk of developing Type II diabetes and where there is limitedadditional capacity for the pancreas further to increase insulinproduction. It would also be inappropriate to increase neutralisingantibody levels to insulin in patients who already have insulin loss asthis may worsen the condition.

It is an aim of the present invention to provide for the intranasaladministration of proteins, such as insulin and insulin analogues, andin particular by the applicant's bi-directional methodology, asdisclosed in the applicant's earlier WO-A-2000/051672, the content ofwhich is incorporated herein by reference.

Delivery of vaccine antigens by the bi-directional methodology has beenshown to date to provide an increase in the circulating antibodies whencompared to intranasal administration by conventional nasal spraytechnologies, in particular nasal spray pumps. As such, a person skilledin the art would have contemplated that intranasal administration ofinsulin using bi-directional delivery technology should increase theimmune response to insulin and other immunogenic proteins, which iscontrary to the requirement for delivery of proteins and peptides to theCNS.

It is a particular aim of the present invention to provide for theintranasal administration of proteins, such as insulin and insulinanalogues, in therapeutically-significant amounts to the olfactory bulbregion in the upper posterior region of the nasal cavity, such that theproteins access the CNS and avoid being presented to the nasalassociated lymphatic tissue (HALT) which includes M-cells, asencompassed by Waldeyer's ring, and in particular in the adenoids, andantigen presenting cells (APCs) which are widely distributed throughoutthe nasal mucosa, which would cause a neutralising antibody response.

The posterior region of the nasal airway is that region which isposterior of the nasal valve NV, as illustrated in FIG. 1. The nasalvalve comprises the anterior bony cavum which contains inferiorturbinate erectile tissue and septal erectile tissue, which aresupported respectively by compliant ala tissue and the rigidcartilaginous septum [10]. These elements combine to form a dynamicvalve, which extends over several millimetres, that adjusts nasalairflow, and is stabilized by cartilage and bone, modulated by voluntarymuscle and regulated by erectile tissue. The lumen of the nasal valve isthe section of narrowest cross-sectional area between the posterior andanterior regions of the nasal airway, and is much longer and narrowerdorsally than ventrally, and this lumen defines a triangular entrancewhich extends to the piriform region of the bony cavum. The nasal valveis lined In its anterior part with transitional epithelium, with agradual transition posterior to respiratory epithelium. The nasal valveand anterior vestibule define roughly the anterior one-third of thenose.

The posterior region of the nasal airway is that region which is linedwith respiratory epithelium, which is ciliated, and olfactoryepithelium, which comprises nerves which extend downwards through thecribiform plate CP from the olfactory bulb OB and defines the olfactorybulb region, whereas the anterior region of the nasal airway is thatregion which is lined with squamous epithelium, which is not ciliated,and transitional epithelium. The olfactory epithelium extends on boththe lateral and medial sides of the nasal airway which defines theolfactory cleft, and typically extends downwards about 1.5 to 2.5 cm.

The upper posterior region is the region above the inferior meatus IM,as illustrated in FIG. 1, and encompasses the middle turbinate, themiddle meatus, the sinus ostia in infundibulum (ostia to maxillary,frontal and ethmoidal sinuses), the olfactory region, and the upperbranches of the trigeminal nerve, and is that region which includesveins which drain to the venous sinuses that surround the brain.

As illustrated in FIG. 1, the posterior region of the nasal airway isthe nasal region posterior of an imaginary vertical plane VERT which islocated at a position corresponding to the lower angle of the anteriornasal aperture (aperture piriformis), which corresponds substantially toone-quarter of the distance between the anterior nasal spine AnS, whichis a pointed projection at the anterior extremity of the intermaxillarysuture, and the posterior nasal spine PnS, which is the sharp posteriorextremity of the nasal crest of the hard palate and represents thetransition between the nose and the nasopharynx, which corresponds to adistance posterior of the anterior nasal spine AnS of between about 13mm and about 14 mm (Rosenberger [11] defines the distance between theanterior nasal spine AnS and the posterior nasal spine PnS as being 56mm in eighteen year old boys and 53.3 mm in eighteen year old girls).

As further illustrated in FIG. 1, the upper region of the nasal airwayis an upper segment of the nasal airway which is bounded by thecribiform plate CP and a horizontal plane HORIZ which is located at aposition corresponding to one-third of the distance between the nasalfloor NF of the nasal airway and the cribiform plate CP, whichcorresponds to a height of typically between about 13 and about 19 mmabove the nasal floor NF (Zacharek et al [12] define the distance fromthe nasal floor NF to the cribiform plate CP as 46+/−4 mm).

The upper posterior region is thus that upper posterior region which isbounded by the above-defined vertical and horizontal planes VERT, HORIZ.

It is a further aim of the present invention to provide for theintranasal administration of proteins, such as insulin and insulinanalogues, such that the proteins which are not absorbed into the CNSremain in solution and are not phagocytosed by immunogenic cells, suchas the antigen presenting cells.

It is a still further aim of the present invention to provide for theintranasal administration of proteins, such as insulin and insulinanalogues, and in particular where delivered as a powdered formulation,such that a minimal amount of associated endotoxin is deliveredtherewith, so as to avoid triggering the maturation of immature antigenpresenting cells.

It is a yet further aim of the present invention to provide that theintranasal administration of insulin, in particular in the treatment ofAlzheimer's disease and obesity, does not result in the exacerbation orprecipitation of type II diabetes.

In one aspect the present invention provides for selective delivery ofimmunogenic proteins to the upper posterior region of the nasal cavity,and in particular the olfactory bulb region, such as to provide foruptake of an immunogenic protein to the CNS without triggering asignificant immune response.

In another aspect the present invention provides for an administrationmethodology which minimizes the delivery of endotoxins.

In one embodiment the nosepiece of the delivery device is configured toprevent the accumulation of endotoxins thereon, such as by way of anagent, for example, as a coating, which degrades polysaccharides, whichrepresent the immunogenic component of endotoxins, for example, by wayof binding to the polysaccharide. In one embodiment a cap for thenosepiece could similarly include such an agent.

In another embodiment the nosepiece of the delivery device isreplaceable. With this methodology, the accumulation of endotoxins onthe nosepiece, as would occur through repeated use, is prevented.

In another aspect the present invention provides a solubilized proteinformulation which is formulated, such as by including one or moresolubilizing agents, to provide for the protein to remain in solutionfollowing intranasal administration.

In this way, the one or more proteins do not precipitate out of solutionfollowing administration, for example, owing to a shift in pH, ionicbalance or osmolarity, which would result in undesirable phagocytosationof the protein by the antigen presenting cells.

In a further aspect the present invention provides for a powderedprotein formulation which provides for the rapid and completedissolution of the protein following administration.

Powdered protein formulations may have advantages in terms of stability,but are likely to result in insoluble particles being phagocytosed bythe antigen presenting cells. The present inventors have recognized thatthis can be overcome by ensuring that the powder particles are rapidlyhydrated following administration.

In a still further aspect the present invention provides for a proteinformulation which includes an immunomodulator, which acts to prevent animmune response to the administered protein.

In a yet still further aspect the present invention provides for aprotein formulation which is formulated to degrade any of the proteinwhich remains to be absorbed within 15 minutes, and preferably 10minutes, following administration.

With this formulation, any of the protein which is transported by themucociliary clearance mechanisms to the nasal associated lymphatictissue and the antigen presenting cells is degraded such that no immuneresponse is raised to the protein.

In yet another aspect the present invention provides a proteinformulation which is formulated to ensure uptake from the olfactory bulbregion within 10 minutes following administration.

In one embodiment the formulation includes an uptake agent, such as acyclodextrin, for providing for rapid uptake of the protein in theolfactory bulb region.

In another embodiment, where the formulation is delivered by thebi-directional methodology, the particle sizes of the deliveredformulation and the flow rate of the entraining gas flow are such as toprovide for targeted delivery to and efficient transfer across theolfactory bulb region, such as to provide for a much reduced antibodyresponse as compared to delivery by conventional nasal spray technology,in particular nasal spray pumps.

With this formulation, none of the protein remains to be transported bythe mucociliary clearance mechanisms to the nasal associated lymphatictissue, including the specialized M-cells, and the antigen presentingcells.

In yet another aspect the present invention provides a delivery devicefor delivering a substance to the nasal airway of a subject, thedelivery device including a nosepiece having a replaceable outer sleeve,with the sleeve preferably including an indicator material whichprovides an indication of exposure to one or more of moisture andbiomaterials, such as endotoxins.

In yet still another aspect the present invention provides a deliverydevice for delivering a substance to the nasal airway of a subject, thedelivery device including a nosepiece having a plurality of removablesleeves which are removable successively to allow for exposure of afresh sleeve, with the sleeves preferably including an indicatormaterial which provides an indication of exposure to one or more ofmoisture and biomaterials, such as endotoxins.

Preferred embodiments of the present invention will now be describedhereinbelow by way of example only with reference to the accompanyingdrawings, in which:

FIG. 1 illustrates the segmentation of a nasal cavity in accordance witha preferred embodiment of the present invention;

FIG. 2 schematically illustrates a nasal delivery device for deliveringa protein formulation to a nasal airway of a subject in accordance withone embodiment of the present invention;

FIG. 3 illustrates the delivery device of FIG. 2 where operative todeliver a dose of the embodied protein formulation into the nasal airwayof the subject;

FIG. 4 illustrates a nasal delivery device as one modification of thefirst-described embodiment, with an outer sleeve of the nosepiecefitted;

FIG. 5 illustrates the delivery device of FIG. 4, with the outer sleeveof the nosepiece removed;

FIG. 6 illustrates a nasal delivery device as another modification ofthe first-described embodiment; and

FIG. 7 illustrates the delivery device of FIG. 6, with a sleeve of thenosepiece removed.

The delivery device comprises a housing 15, a nosepiece unit 17 forfitting in a nasal cavity of a subject, and a mouthpiece 19 throughwhich the subject exhales to actuate the delivery device.

The nosepiece unit 17 comprises a nosepiece 20, in this embodiment afrusto-conical element, for guiding the nosepiece unit 17 into a nasalpassage of the subject and being configured both to provide afluid-tight seal with the nares of the nostril and obstruct, in thisembodiment close, the nasal passage at a position therealong, in thisembodiment at a position corresponding substantially to the nasal valve,thereby obstructing the anterior one-third of the nasal passage andleaving open the posterior two-thirds of the nasal passage, asillustrated in FIG. 3, and an outlet unit 21 for delivering substance,in this embodiment a protein formulation containing insulin, to an upperposterior region of the nasal passage of the subject, in this embodimentan upper posterior region as bounded by a vertical plane which islocated posterior of the anterior nasal spine AnS at a positioncorresponding to one-quarter of the distance between the anterior andposterior nasal spines AnS, PnS and a horizontal plane which is locatedabove the nasal floor at a height one-third of the distance between thenasal floor and the cribiform plate, which encompasses the olfactorybulb region from which the substance is uptaken into the CNS of thesubject.

In this embodiment the delivery device is re-usable, and the nosepiece20 is replaceable, such as to prevent the accumulation of endotoxinsthereon, which could be transferred to the nasal cavity of the subject.

In an alternative embodiment the nosepiece 20 could be configured toprevent accumulation of endotoxins thereon, such as by including ananti-bacterial.

In this embodiment the outlet unit 21 comprises a delivery channel 23which is in fluid communication with the mouthpiece 19 such that an airflow is delivered into and through the nasal airway of the subject onexhalation by the subject through the mouthpiece 19, and a nozzle 25 fordelivering the nasal formulation to the nasal airway of the subject.

In this embodiment the nozzle 25 is configured to deliver an aerosolspray, either as a liquid or a powder aerosol spray, but in analternative embodiment the nozzle could be configured to deliver a jet,that is, as a column of the formulation, either as a liquid or powderjet.

The delivery device further comprises a substance supply unit 29 fordelivering metered doses of the formulation, which is fluidly connectedto the nozzle 25 to deliver the nasal formulation from the nosepiece 17,in this embodiment as an aerosol spray.

In this embodiment the substance supply unit 29 comprises a mechanicaldelivery pump, in particular a liquid delivery pump or a powder deliverypump, which delivers metered doses of substance, on actuation thereof.

In another alternative embodiment the substance supply unit 29 couldcomprise a dry powder delivery unit which delivers metered doses ofsubstance, as a dry powder, on actuation thereof. In one embodiment thesubstance supply unit 29 could provide for delivery of substance from acapsule.

In yet another alternative embodiment the substance supply unit 29 couldcomprise an aerosol canister which delivers metered volumes of apropellant, preferably a hydrofluoroalkane (HFA) propellant or the like,containing substance, either as a suspension or solution.

In this embodiment the substance supply unit 29 is a multi-dose unit fordelivering a plurality of metered doses of the nasal formulation. Inanother embodiment the substance supply unit 29 could be a single-doseunit for delivering a single metered dose of the nasal formulation.

The substance supply unit 29 is pre-primeable, in this embodiment byloading a resilient element, and includes a breath-actuated releasemechanism 31 which, when triggered, releases the resilient element andactuates the substance supply unit 29 to deliver a metered dose of thenasal formulation through the nozzle 25.

In this embodiment the trigger mechanism 31 is configured to causeactuation of the substance supply unit 29 on generation of apredetermined flow rate through the delivery channel 23.

In one embodiment the protein formulation comprises a solubilizedprotein formulation.

In one embodiment the solution comprises a viscous solution, such as agel.

In one embodiment the protein formulation is such as to remain insolution following delivery, and preferably includes a solubilizingagent for maintaining the protein in solution following delivery.

In one embodiment the protein formulation is such that the protein doesnot precipitate from solution owing to one or more of a shift in pH,ionic balance or osmolarity following delivery.

In an alternative embodiment the protein formulation could be a powderedprotein formulation.

In one embodiment the powdered protein formulation provides fordissolution of the protein following delivery.

Preferably, the powdered protein formulation provides for dissolution ofthe protein within about 5 minutes of delivery.

More preferably, the powdered protein formulation provides fordissolution of the protein within about 2 minutes of delivery.

Still more preferably, the protein formulation provides for dissolutionof the protein within about 1 minute of delivery.

In one embodiment the protein formulation includes an immunomodulator,which acts to prevent an immune response to the protein.

In one embodiment the protein formulation is such as to degrade theprotein which remains to be absorbed subsequent to a predeterminableperiod following delivery.

In one embodiment the protein formulation includes a proteolytic agentwhich acts to degrade the protein which remains to be absorbedsubsequent to the predeterminable period following delivery.

In one embodiment the proteolytic agent can include one or more oftrypsin, chymotrypsin, N terminal peptidases and C terminal peptidases.

Preferably, the protein formulation is such as to degrade the proteinwhich remains to be absorbed within about 15 minutes following delivery.

More preferably, the protein formulation is such as to degrade theprotein which remains to be absorbed within about 10 minutes followingdelivery.

Still more preferably, the protein formulation is such as to degrade theprotein which remains to be absorbed within about 5 minutes followingdelivery.

In one embodiment the protein formulation provides for rapid uptake ofthe protein from the olfactory bulb region in the upper posteriorregion.

In one embodiment the protein formulation includes an uptake agent forproviding for uptake of the protein from the olfactory bulb region.

In one embodiment the uptake agent is a cyclodextrin.

Preferably, the protein formulation provides for uptake of the proteinfrom the olfactory bulb region within about 10 minutes followingdelivery.

More preferably, the protein formulation provides for uptake from theolfactory bulb region within about 5 minutes following delivery.

In an alternative embodiment the protein formulation comprises anantidiuretic hormone, such as argipressin, lypressin, desmopressin,felypressin, ornipressin, terlipressin and vasopressin or theirpharmaceutically-acceptable derivatives or analogues.

In an alternative embodiment the protein formulation comprises anoxytocic hormone, such as carbetocin, demoxytocin and oxytocin or theirpharmaceutically-acceptable derivatives or analogues.

In an alternative embodiment the protein formulation comprises anoxytocin antagonist, such as atosiban or its pharmaceutically-acceptablederivatives or analogues.

In an alternative embodiment the protein formulation comprises acorticotrophic hormone, such as corticotrophin and tetracosactide ortheir pharmaceutically-acceptable derivatives or analogues.

In an alternative embodiment the protein formulation comprises acorticotrophic releasing hormone, such as corticorelin or itspharmaceutically-acceptable derivatives or analogues.

In an alternative embodiment the protein formulation comprises anomatotrophic hormone, such as mecasermin, somatrem and somatropin ortheir pharmaceutically-acceptable derivatives or analogues.

In an alternative embodiment the protein formulation comprises asomatotrophic hormone receptor antagonist, such as pegvisomant or itspharmaceutically-acceptable derivatives or analogues.

In an alternative embodiment the protein formulation comprises anomatotrophic releasing hormone, such as sermorelin and somatorelin ortheir pharmaceutically-acceptable derivatives or analogues.

In an alternative embodiment the protein formulation comprises asomatotrophic release inhibitor, such as lanreotide, octreotide,somatostatin and vapreotide or their pharmaceutically-acceptablederivatives or analogues.

In an alternative embodiment the protein formulation comprises agonadotrophic hormone, such as choriogonadotrophin alfa, chorionicgonadotrophin, a follicle stimulating hormone, follitropin alfa,follitropin beta, a luteinising hormone, lutropin alfa, menotrophin andurofollitropin or their pharmaceutically-acceptable derivatives oranalogues.

In an alternative embodiment the protein formulation comprises agonadotrophic releasing hormone, such as buserelin, deslorelin,gonadorelin, goserelin, histrelin, leuprorelin, naferlin and triptorelinor their pharmaceutically-acceptable derivatives or analogues.

In an alternative embodiment the protein formulation comprises anonadotrophic releasing hormone antagonist, such as abarelix, cetorelixand ganirelix or their pharmaceutically-acceptable derivatives oranalogues.

In an alternative embodiment the protein formulation comprises athyrotrophic hormone, such as thyrotrophin and thyrotrophin alfa ortheir pharmaceutically-acceptable derivatives or analogues.

In an alternative embodiment the protein formulation comprises athyrotrophic releasing hormone, such as posatirelin, protirelin andtaltirelin or their pharmaceutically-acceptable derivatives oranalogues.

In an alternative embodiment the protein formulation comprises alactotrophic hormone, such as prolactin or itspharmaceutically-acceptable derivatives or analogues.

In an alternative embodiment the protein formulation comprises ametabolic peptide, such as an insulin-like growth factor, a glucagon, agrowth hormone and PYY3-36 or their pharmaceutically-acceptablederivatives or analogues.

In an alternative embodiment the protein formulation comprisescalcitonin or pharmaceutically-acceptable derivatives or analoguesthereof, such as elcatonin and salcatonin.

In an alternative embodiment the protein formulation comprises amelanocyte stimulating hormone.

In an alternative embodiment the protein formulation comprises a nervegrowth factor.

In an alternative embodiment the protein formulation comprises anepidermal growth factor.

In an alternative embodiment the protein formulation comprises anepoetin or its pharmaceutically-acceptable derivatives or analogues.

In an alternative embodiment the protein formulation comprises aninterleukin.

In an alternative embodiment the protein formulation comprises a proteininvolved in one or both of blood coagulation and fibrinolysis.

In an alternative embodiment the protein formulation comprises anantibiotic.

Operation of the delivery device will now be described hereinbelow withreference to FIG. 3 of the accompanying drawings.

The nosepiece 17 is first inserted into one of the nasal cavities of asubject until the nosepiece 20 abuts the nares of the nostril, at whichpoint the distal end of the outlet unit 21 extends about 2 cm into thenasal cavity of the subject, and the mouthpiece 19 is gripped in thelips of the subject.

The subject then begins to exhale through the mouthpiece 19, whichexhalation acts to close the oropharyngeal velum of the subject anddrive an air flow through the delivery channel 23 of the outlet unit 21,with the air flow passing into the one nasal cavity, around theposterior margin of the nasal septum and out of the other nasal cavity,thereby achieving a bi-directional air flow through the nasal airway ofthe subject.

In this embodiment, when the flow rate developed through the deliverychannel 23 reaches a predetermined value, the release mechanism 31 istriggered to actuate the substance supply unit 29 to deliver a metereddose of the nasal formulation to the nozzle 25 and into the nasal cavityof the subject as an aerosol spray.

In this embodiment, where the delivery device is a multi-dose device,the device is ready for further use following priming of the substancesupply unit 29.

Finally, it will be understood that the present invention has beendescribed in its preferred embodiments and can be modified in manydifferent ways without departing from the scope of the invention asdefined by the appended claims.

In one modification of the above-described device, as illustrated inFIGS. 4 and 5, the nosepiece 20 could include a replaceable outer sleeve35, which allows for replacement at periodic intervals, for example,after each operation. FIGS. 4 and 5 illustrate the sleeve 35 when fittedand removed, respectively.

In one embodiment the sleeve 35 can include an Indicator material whichprovides an indication of exposure to one or more of moisture andbiomaterials, for example, endotoxins, thereby providing an indicationto the user when the sleeve 35 should be replaced.

In another modification of the above-described device, as illustrated inFIGS. 6 and 7, the nosepiece 20 could include a plurality of removablesleeves 37, typically formed as thin films, which are peelablesuccessively to allow for exposure of a fresh sleeve 37 at periodicintervals, for example, after each operation.

In one embodiment one or more of the innermost sleeves 37 can be marked,for example, colored, to indicate that a minimum number of the sleeves37 remain.

In one embodiment the sleeves 37 can include an indicator material whichprovides an indication of exposure to one or more of moisture andbiomaterials, for example, endotoxins, thereby providing an indicationto the user when the outer sleeve 37 should be discarded.

Furthermore, in the above-described embodiment the delivery device isconfigured to deliver an air flow through one nostril of a subject atsuch a pressure as to flow around the posterior margin of the nasalseptum and out of the other nostril of the subject, thereby achievingbi-directional delivery through the nasal cavities as disclosed inWO-A-2000/51672, the content of which is herein incorporated byreference, but in an alternative embodiment the delivery device could beconfigured to deliver an air flow which is not sufficient to achievebi-directional delivery through the nasal cavities or utilizes noentraining gas flow. This embodiment is still advantageous as comparedto known delivery devices, in providing for velum closure and beingcapable of achieving targeted delivery, particularly when certainregions of the nasal cavity are obstructed by cuff members.

In another alternative embodiment the above-described delivery devicecould be configured not to provide for any gas flow, but instead providefor targeted delivery through use of an optimized nosepiece.

REFERENCES

-   1. Baker, L D et al, Acute intranasal insulin administration    improves verbal memory for adults with Alzheimer's disease, Society    for Neuroscience Abstract Viewer and Itinerary Planner 2003,    Abstract No 84.17.-   2. Benedict, C et al, Intranasal insulin improves memory in humans,    Psychoneuroendocrinology, November 2004, 29, pages 1326-34.-   3. Born, J et al, Sniffing neuropeptides: a transnasal approach to    the human brain, Nat Neurosci, June 2002, 5(6), pages 514-6.-   4. Fehm, L et al, Body weight regulation through the central nervous    system. The development of a pathogenetically based adiposity    therapy, Med Klin (Munich), November 2004, 99(11), pages 674-9.-   5. Hallschmid, M et al, Intranasal insulin reduces body fat in men    but not in women, Diabetes, November 2004, 53, pages 3024-9.-   6. Hallschmid, M et al, Manipulating central nervous mechanisms of    food intake and body weight regulation by intranasal administration    of neuropeptides in man, Physiol Behav, October 2004, 83, pages    55-64.-   7. Stockhorst, U et al, Insulin and the CNS: effects on food intake,    memory, and endocrine parameters and the role of intranasal insulin    administration in humans, Physiol Behav, October 2004, 83, pages    47-54.-   8. Watson, G S et al, Insulin effects on CSF norepinephrine and    cognition in Alzheimer's disease, Neurobiology of Aging, January    2006, 27(1), pages 38 to 41.-   9. Harrison, L C et al, Administration of Intranasal Insulin to    Humans At Risk for Type I Diabetes, Diabetes Care, 2004, 27 pages    2348-2355.-   10. Cole, P, The Respiratory Role of the Upper Airways, a selective    clinical and pathophysiological review. 1993, Mosby-Year Book Inc.    ISBN1.55664-390-X.-   11. Rosenberger, H, Growth and Development of the Naso-Respiratory    Area in Childhood, PhD Thesis, Laboratory of Anatomy, School of    Medicine, Western Reserve University, Presented to the Annual    Meeting of the American Laryngological, Rhinological and Otological    Society, Charleston, S.C., USA, 1934.-   12. Zacharek, M A et al, Sagittal and Coronal Dimensions of the    Ethmoid Roof: A Radioanatomic Study, Am J Rhinol 2005, 19, pages    348-352.-   13. Bakke H et al, Oral Spray Immunization May Be an Alternative to    Intranasal Vaccine Delivery to Induce Systemic Antibodies but not    Nasal Mucosal or Cellular Immunity, Scandinavian Journal of    Immunology, March 2006, 63, pages 223-31.-   14. Brandtzæg P et al, Role of secretory antibodies in the defence    against infections, Int J Med Microbiol, April 2003, 293(1), pages    3-15.

For the avoidance of doubt, the contents of the above-identifiedreferences are herein incorporated by reference.

1. A delivery device for delivering a protein formulation to the upperposterior region of a nasal cavity of a subject for uptake into the CNSof the subject, the delivery device comprising a nosepiece unitincluding a nosepiece for fitting to a nostril of a subject and a nozzlethrough which the formulation is in use delivered to the respectivenasal cavity, and a delivery unit for delivering formulation through thenozzle of the nosepiece, wherein the delivery of the formulation is suchas to provide for uptake of the protein to the CNS without triggering asignificant immune response. 2-55. (canceled)
 56. A method of deliveringa protein formulation to the upper posterior region of a nasal cavity ofa subject for uptake into the CNS of the subject, the method comprisingthe step of delivering the formulation through a nozzle of a nosepieceof a delivery device into the nasal cavity of the subject, such as toprovide for uptake of the protein to the CNS without triggering asignificant immune response. 57-110. (canceled)
 111. A proteinformulation for delivery to a nasal cavity of a subject for uptake intothe CNS of the subject, the formulation being such as to provide foruptake of the protein to the CNS without triggering a significant immuneresponse. 112-158. (canceled)
 159. A delivery device for delivering asubstance to the nasal airway of a subject, the delivery deviceincluding a nosepiece having a replaceable outer sleeve.
 160. Thedelivery device of claim 159, wherein the sleeve includes an indicatormaterial which provides an indication of exposure to one or more ofmoisture and biomaterials, such as endotoxins.
 161. A delivery devicefor delivering a substance to the nasal airway of a subject, thedelivery device including a nosepiece having a plurality of removablesleeves which are removable successively to allow for exposure of afresh sleeve.
 162. The delivery device of claim 161, wherein the sleevesare individually peelable films.
 163. The delivery device of claim 161,wherein one or more of the innermost sleeves are marked to indicate thata predeterminable number of the sleeves remain.
 164. The delivery deviceof claim 161, wherein the sleeves include an indicator material whichprovides an indication of exposure to one or more of moisture andbiomaterials, such as endotoxins.